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Extra kinetic dimensions for label discrimination

Author

Listed:
  • Raja Chouket

    (PSL University, Sorbonne Université, CNRS)

  • Agnès Pellissier-Tanon

    (PSL University, Sorbonne Université, CNRS)

  • Aliénor Lahlou

    (PSL University, Sorbonne Université, CNRS
    Sony Computer Science Laboratories)

  • Ruikang Zhang

    (PSL University, Sorbonne Université, CNRS)

  • Diana Kim

    (PSL University, Sorbonne Université, CNRS)

  • Marie-Aude Plamont

    (PSL University, Sorbonne Université, CNRS)

  • Mingshu Zhang

    (Chinese Academy of Sciences)

  • Xi Zhang

    (Chinese Academy of Sciences)

  • Pingyong Xu

    (Chinese Academy of Sciences
    University of Chinese Academy of Sciences)

  • Nicolas Desprat

    (PSL University, CNRS, Sorbonne Université, Université de Paris
    PSL University)

  • Dominique Bourgeois

    (Univ. Grenoble Alpes, CNRS, CEA, IBS)

  • Agathe Espagne

    (PSL University, Sorbonne Université, CNRS)

  • Annie Lemarchand

    (LPTMC, Sorbonne Université, CNRS)

  • Thomas Le Saux

    (PSL University, Sorbonne Université, CNRS)

  • Ludovic Jullien

    (PSL University, Sorbonne Université, CNRS)

Abstract

Due to its sensitivity and versatility, fluorescence is widely used to detect specifically labeled biomolecules. However, fluorescence is currently limited by label discrimination, which suffers from the broad full width of the absorption/emission bands and the narrow lifetime distribution of the bright fluorophores. We overcome this limitation by introducing extra kinetic dimensions through illuminations of reversibly photoswitchable fluorophores (RSFs) at different light intensities. In this expanded space, each RSF is characterized by a chromatic aberration-free kinetic fingerprint of photochemical reactivity, which can be recovered with limited hardware, excellent photon budget, and minimal data processing. This fingerprint was used to identify and discriminate up to 20 among 22 spectrally similar reversibly photoswitchable fluorescent proteins (RSFPs) in less than 1s. This strategy opens promising perspectives for expanding the multiplexing capabilities of fluorescence imaging.

Suggested Citation

  • Raja Chouket & Agnès Pellissier-Tanon & Aliénor Lahlou & Ruikang Zhang & Diana Kim & Marie-Aude Plamont & Mingshu Zhang & Xi Zhang & Pingyong Xu & Nicolas Desprat & Dominique Bourgeois & Agathe Espagn, 2022. "Extra kinetic dimensions for label discrimination," Nature Communications, Nature, vol. 13(1), pages 1-8, December.
    • Handle: RePEc:nat:natcom:v:13:y:2022:i:1:d:10.1038_s41467-022-29172-0
    DOI: 10.1038/s41467-022-29172-0
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    References listed on IDEAS

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